一种简便快速的液相色谱串联质谱法用于人体内血尿样品中奈诺沙星浓度的测定  

A rapid and robust method for determination of nemonoxacin in human plasma and urine

作　　者：黄俊[1] 郭蓓宁[1] 郁继诚[1] 武晓捷[1] 曹国英[1] 张菁[1] 施耀国[1] 张婴元[1] 

机构地区：[1]复旦大学附属华山医院抗生素研究所 卫生部抗生素临床药理重点实验室,上海200040

出　　处：《中国感染与化疗杂志》2012年第4期313-318,共6页Chinese Journal of Infection and Chemotherapy

基　　金：国家科技部"重大新药创制"科技重大专项资助项目(2012ZX09303004-001)

摘　　要：目的建立一种简便快速高效液相色谱串联质谱(LC-MS/MS)的方法用于测定人血浆和尿液中奈诺沙星药物浓度。方法色谱柱为Waters Symmetry RP1 8柱(50mm×2.1 mmm,5μm),以加替沙星为内标,血浆和尿液样品测定采用0.1%甲酸乙腈混合溶液为流动相,比例分别为82:1 8和85:1 5,流速均为0.2 mL/min。采用蛋白沉淀法处理血浆样品,以液液萃取法清除尿液样品中杂质。ESI源正离子选择性反应临测(SRM),奈诺沙星检测通道:m/z 372.4→m/z354.1,内标检测通道:m/z376.1→m/z 332.2。结果本方法测定血浆和尿液中的奈诺沙星的线性范围均为5～100ng/mL,最低检测浓度均为5ng/mL血浆和尿液的奈诺沙星提取回收率分别为(98.49±4.69)%和(84.92±6.81)%,且血浆和尿液的RSD)分别小于或等于4.31%和9.76%。奈诺沙星血浆样品的方法日内、日间准确度分别为(99.27～103.75)%和(100.40～106.36)%;而尿液样品的方法日内、日间准确度依次为(103.52～11 5.94)%和(9 9.50～1 10.96)%,且基质效应、精密度、稳定性等均通过方法学验证。并已用于该药的人体药动学研究中药物浓度的测定。结论本方法操作简便、迅速,特异性强,灵敏度高,准确性好,符合奈诺沙星人体药物动力学研究中血尿样测定的方法学要求。Objective A rapid, sensitive and selective liquid chromatography-tandem mass spectrometric （LC-MS/MS） method for the quantification of nemonoxacin in human plasma and urine was developed and validated. Methods The plasma and urine samples were subjected to acetonitrile protein precipitation and liquid-liquid extraction, respectively. Chromatographic separa- tion was performed on a C18 column using an isocratic elution. The mobile phase consisted of 0.1% of formic acid in water and acetonitrile and was pumped at a flow rate of 0. 2 mL/min. Quantitative analysis was conducted in the selected reaction monito- ring mode with preeursor/produet ion transitions of m/z 372.4/354. 1 and m/z 376. 1/332.2, for nemonoxacin and internal standard respectively. Results The calibration eurves were linear （r= 0. 9994 for plasma, r = 0. 9989 for urine, respectively） within the range of 5-1 000 ng/mL and the lower limit of quantification was 5 ng/rnL for both. The recovery of nemonoxaein was （98.49 ± 4.69）% for plasma and （84.92± 6.81）% for urine samples, respectively. The inter-day and intra-day relative standard deviations （RSDs） of nemonoxacin plasma and urine samples were less than, if not, equal to 4.31% and 9.76%, re- spectively. The inter-day and intra-day accuracy of nemonoxacin plasma samples was （99. 27-103. 75）% and （100. 40- 106.36）%; while the inter-day and intra-day accuracy of nemonoxacin urine samples was （103. 52-115.94）%and （99. 50- 110.96） %, respectively. The matrix effect, precision, selectivity and stability were validated for nemonoxacin both in human plasma and urine. Conclusions This method is simple, rapid, specific and precise for the determination of nemonoxacin in hu- man plasma and urine. It is applicable for pharmacokinetic study of nemonoxacin.

关 键 词：奈诺沙星 液相色谱串联质谱法 方法学验证 应用 

分 类 号：R96[医药卫生—药理学]